Germ cell apoptosis and expression of proteins related to cell apoptosis in zebrafish testes exposed to DBDPE in vivo. (A) Apoptosis detection in zebrafish testes by immunofluorescence staining using TUNEL assay. The representative images show DAPI-stained (blue) nuclei with TUNEL-positive signals in green. Scale bar: 20μm. (B) Statistical analysis of the total number of TUNEL-positive cells relative to the section areas. Each dot in (B) represents one replicate data point (mean value of each testis). The dot numbers represent the data size (n=8–12) for statistical analysis. (C) Western blotting analyses using antibodies against cleaved caspase-3, cleaved PARP, p-JNK, and GAPDH in testes tissues. The numbers on the left represent molecular weight. (D) Quantification of the abundances of proteins relative to GAPDH (n=3). Results are represented as means±standard errors of the mean (SEMs). Data are reported in Excel Table S6. Note: DAPI, 4′,6-diamidino-2′-phenylindole; DBDPE, decabromodiphenyl ethane; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PARP, poly(adenosine diphosphosphate-ribose) polymerase; p-JNK, phospho-c-jun N-terminal kinase; TUNEL, terminal deoxynucleotidyl transferase deoxynucleotide triphosphate nick-end labeling (assay). *p<0.05, **p<0.01, and ***p<0.001 indicate significant differences between exposure and control groups, by one-way analysis of variance (ANOVA) followed by the post hoc least significant difference (LSD) test.
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